Immunohistochemical and Immunofluorescent Analysis of Lung Tissues

Formalin-fixed and paraffin-embedded lung tissues were cut into sections of 3 um in thickness for immunohistochemistry (IHC). 4% paraformaldehyde-fixed lung tissues were cut into sections of 6 um in thickness for immunofluorescence (IF) staining. IHC and IF were performed as previously (31 (link)). The primary antibody used were SDF4 (1:100 dilution in 3% BSA), ATF6 (1:200 dilution in 3% BSA), GRP78 (1:200 dilution in 3% BSA) and CHOP (1:200 dilution in 3% BSA). Images were acquired using Leica DM4000 microscope and Nikon A1 Ti confocal microscope.

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Zhu T., Su Q., Wang C., Shen L., Chen H., Feng S., Peng X., Chen S., Wang Y., Jiang H, & Chen J. (2021). SDF4 Is a Prognostic Factor for 28-Days Mortality in Patients With Sepsis via Negatively Regulating ER Stress. Frontiers in Immunology, 12, 659193.

Publication 2021

DM4000 microscope A1 Ti confocal microscope

Antibody Atf6 Chop Confocal microscope Dilution Formalin Grp78 Immunofluorescence Immunohistochemistry Lung Microscope Paraffin Paraformaldehyde Tissues

Corresponding Organization : Zhejiang University

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Variable analysis

independent variables

Formalin-fixed and paraffin-embedded lung tissues
4% paraformaldehyde-fixed lung tissues

dependent variables

Immunohistochemistry (IHC) staining
Immunofluorescence (IF) staining

control variables

Thickness of tissue sections (3 um for IHC, 6 um for IF)
Primary antibody dilutions (SDF4 1:100, ATF6 1:200, GRP78 1:200, CHOP 1:200 in 3% BSA)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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